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Artificial blood vessels carrying adenovirus-mediated vascular endothelial growth factor gene in endothelial cell growth

Author: JiangDaMing
Tutor: LiuChangWei；WangLinFang
School: Peking Union Medical College , China
Course: Clinical
Keywords: Artificial blood vessels Vascular endothelial growth factor Endothelial cells Endothelialization Adenovirus
CLC: R318.1
Type: PhD thesis
Year: 2008
Downloads: 54
Quote: 1
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Abstract


Background: The small diameter artificial vascular graft intimal hyperplasia and postoperative thrombosis, resulting in patency rate is not satisfactory, mainly because of the lack of complete artificial blood vessels within the cortical surface. It has been confirmed through rapid endothelialization of vascular prosthesis surface can reduce the incidence of restenosis. Artificial vascular endothelium of the research has progressed to the endothelial cells from growing genetic and molecular level. Vascular endothelial growth factor (VEGF) induces vascular endothelial cell growth is strongest, one of the highest specific growth factor. VEGF protein and gene plasmid coating has its obvious flaws. The adenovirus as a vector transfection achieved good results. Objective: To observe the artificial blood vessels that carry adenovirus-mediated endothelial cell VEGF gene transfection effect, VEGF gene and protein expression as well as on endothelial cell growth. Materials and Methods: 1.ad5-EGFP transfected EAhy926 transfection rate determination. Two artificial blood vessels that carry adenovirus release in still conditions. Three artificial vascular endothelial cell adhesion on the situation. 4.ad5-VEGF165-IRES-EGFP transfected EAhy926, RT-PCR semi-quantitative detection of VEGF gene expression levels. 5 artificial blood vessels that carry ad5-VEGF165-IRES-EGFP transfected EAhy926, ELISA assay levels of VEGF protein expression. 6 artificial blood vessels that carry ad5-VEGF165-IRES-EGFP transfected EAhy926, MTT assay on endothelial cell growth. Results: 1.ad5 transfection efficiency increased with the MOI, MOI 200 reached 87.3%. Two artificial blood vessels digested fluorescent cells in the initial 1h, the decline is most obvious. Basically no dilution of fluorescing cells. 3 electron microscopy showed that endothelial cells attached to the artificial blood vessels. 4.ad5-VEGF165 transfected with VEGF gene RT-PCR expression, the first three days than the first one day expressed strong. The control group did not express. 5.ad5-VEGF165 transfected with supernatant VEGF protein, showing an upward trend in the first five days after the peak began to decline. The control group did not express. 6.ad5-VEGF165 transfected group OD values ??rose rapidly in the control group increased slowly. Conclusion: 1.ad5 optimal transfection EAhy926 MOI of 200.2. Artificial blood vessels carry the virus still water retention in the artificial blood vessel about 1h, which is a gradual release process, rather than the instantaneous release. 3 endothelial cells can be attached in an artificial blood vessels. 4.ad5-VEGF165-IRES-EGFP can be transfected EAhy926, hVEGF165 gene expression levels of success. 5 artificial blood vessels that carry ad5-VEGF165-IRES-EGFP could transfection EAhy926, hVEGF165 protein expression levels of success 6. Artificial blood vessels that carry ad5-VEGF165-IRES-EGFP can promote endothelial cell proliferation.

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CLC: > Medicine, health > Basic Medical > Medical science in general > Biomedical Engineering > Artificial organs and organ
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